Investigating the mechanism of action of Bu-Yang-Huan-Wu decoction in treating bleomycin-Induced pulmonary fibrosis through the epithelial-mesenchymal transition pathway

Background:To explore the effects and mechanisms of Bu-Yang-Huan-Wu Decoction on pulmonary fibrosis in mice.Methods:Forty-five C57BL/6J mice were randomly divided into three groups:Control,Model,and Bu-Yang-Huan-Wu Decoction.Pulmonary fibrosis was elicited in mice through a solitary intratracheal administration of 2.5 mg/kg bleomycin.For the control group,mice were given a solitary intratracheal administration of a comparable volume of PBS.Treatment began on the first day after the successful model establishment and lasted for 21 days.The survival rate and body weight of the mice were recorded daily,and on the 22nd day,bronchoalveolar lavage fluid was collected to determine total cells and total protein.The wet/dry weight ratio of lung tissue and hydroxyproline were measured.Lung tissue pathology was observed using hematoxylin and eosin staining and Masson staining.The mRNA expression of epithelial-mesenchymal transition-related proteins(E-cadherin and vimentin)was detected by RT-qPCR,and their protein expression was analyzed by western blot.Results:Compared to the model group,the Bu-Yang-Huan-Wu Decoction treatment notably enhanced both the survival rate and body weight in pulmonary fibrosis mice,significantly reduced lung tissue wet/dry weight ratio,total cells,and protein in bronchoalveolar lavage fluid,and hydroxyproline content.The pathological morphology of lung tissue was significantly improved,with increased expression of the epithelial cell marker E-cadherin mRNA and protein,and decreased expression of the mesenchymal cell marker vimentin mRNA and protein.Conclusion:Bu-Yang-Huan-Wu Decoction can improve the degree of bleomycin-induced pulmonary fibrosis in mice by inhibiting epithelial-mesenchymal transition.

Modulatory effect of D-pinitol on bleomycin-induced pulmonary fibrosis in rats

Objective:To assess the effect of D-pinitol on pulmonary fibrosis induced by bleomycin.Methods:Sprague-Dawley rats received intratracheal bleomycin(6 IU/kg)to induce pulmonary fibrosis,followed by administration of either D-pinitol(5,10,or 20 mg/kg)or vehicle or methylprednisolone(10 mg/kg)over 28 days after bleomycin administration.Lung function,biochemical parameters,serum biochemistry,mRNA expressions,and histological features were observed.Results:D-pinitol at 10 and 20 mg/kg significantly(P<0.05)attenuated bleomycin-induced bronchoalveolar lavage fluid,decreased myeloperoxidase,nitric oxide,malondialdehyde levels,and increased glutathione and superoxide dismutase level.D-pinitol also improved lung function(enhanced pause,frequency of breathing,expired volume,and tidal volume).Besides,D-pinitol significantly(P<0.05)upregulated Nrf2 and downregulated mRNA expressions of TGF-β,collagen-1,and Smad-3.Furthermore,considerably less inflammation(peribronchial,perivascular,and total),Ashcroft,and interstitial fibrosis scores were observed in the D-pinitol group.Conclusions:D-pinitol exerts its effect against bleomycin-induced pulmonary fibrosis via antioxidative and anti-fibrotic pathways.

Pharmacodynamic study of cannabidiol on bleomycin-induced pulmonary fibrosis in rats

Objective:To study the protective effect of cannabidiol(CBD)on rats with pulmonary fibrosis and explore the possible mechanism of the use of CBD in the treatment of pulmonary fibrosis.Methods:Sixty SD rats were randomly divided into the normal control group,model group,prednisone group,CBD low,medium and high dose groups(12,36,108 mg/kg,ig),10 rats in each group.Except for the normal control group,the other 5 groups were all induced by tracheal injection of bleomycin to rat models of pulmonary fibrosis.After modeling,the rats were given intragastric administration once a day for 28 consecutive days and samples were taken.The degree of pulmonary edema was detected;the pathological changes of lung tissue were observed by HE and Masson staining;tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)and lung tissue superoxide dismutase(SOD),malondialdehyde(MDA),hydroxyproline(HYP)contents were measured by ELISA,transforming growth factor-β1(TGF-β1)andα-smooth muscle protein(α-SMA)concentration were detected by immunocytochemical method,real-time fluorescent quantitative PCR(qRT-PCR)method was used to detect the mRNA expression levels of TGF-β1,α-SMA,Nrf2 and nuclear transcription factor-κB p65(NF-κB p65).Results:The lung organ coefficient and W/D value were significantly decreased in the CBD administration group(P<0.05);medium and high doses of CBD could reduce the number of collagen fibers and fibroblasts;the pulmonary fibrosis in the low,medium,and high dose groups of CBD was significantly lower.The levels of TNF-α,IL-1β,and IL-6 in rat serum,as well as MDA and HYP in lung tissue,were significantly lower compared to the model group.Additionally,the level of SOD was significantly increased(P<0.05);The expression ofα-SMA was decreased compared with the model group(P<0.05);the contents of TGF-β1,α-SMA and NF-κB p65 mRNA in lung tissue decreased,and the expression level of Nrf2 mRNA increased(P<0.05).Especially,the high-dose group had the most significant effect.Conclusion:CBD can significantly reduce the degree of pulmonary fibrosis in rats,and its potential mechanism may be related to inhibiting inflammatory response,enhancing antioxidant capacity and inhibiting the protein expression of TGF-β1 andα-SMA.

Assessment of traditional Chinese medicine pattern in a bleomycininduced pulmonary fibrosis mouse model: A pilot study

Objective:To initially explore traditional Chinese medicine patterns in a bleomycin-induced pulmonary fibrosis mouse model.Methods:Thirty-six C57BL/6 mice were divided by the random number table method(with 12 rats per group)into three groups:a blank group,a model group,and a number 2 Feibi recipe(FBR-2)group.The pulmonary fibrosis mouse model was established by intratracheal instillation of bleomycin.The FBR-2 group was treated with FBR-2 for 4 weeks.Symptoms in the mice such as mental behavior,food/water intake,body weight,body temperature,respiratory rate,and tongue image were observed.The samples were collected on the 14th day and 28th day after modeling,and lung tissues were visually assessed and microscopically evaluated by staining with hematoxylin-eosin and Masson.The expression levels of hydroxyproline,interleukin(IL)-33,IL-37,tissue plasminogen activator,and plasminogen activator inhibitor-1 were determined by enzyme-linked immunosorbent assay.Results:Mice in the model group were poor in spirit,less active,slow in response,showed reduced food/water intake,body temperature,and body weight,increased respiratory rate,and their tongue color had changed from light red to dark red.However,treatment with FBR-2 significantly improved these symptoms.Extensive inflammatory cell infiltration and collagen fiber deposition were observed in the lung tissues of the model group.Compared with the blank group,the levels of hydroxyproline,IL-33,and plasminogen activator inhibitor-1 in the model group significantly increased(all P<.05),whereas that of tissue plasminogen activator significantly decreased on the 14th day and 28th day(P=.036 and P=.005,respectively).Moreover,FBR-2 improved lung inflammation and fibrinolysis imbalance and reduced collagen fiber deposition.Conclusion:To some extent,our bleomycin-induced pulmonary fibrosis mouse model exhibited traditional Chinese medicine patterns of qi deficiency,blood stasis,and heat retention.

Experimental Study on Effect of Compound Biejia Ruangan Prescription (复方鳖甲软肝方) on High Resolution Computerized Tomographic Images in Bleomycin Induced Pulmonary Fibrosis Rats

Objective: To study the therapeutic effect of Com pound Biejia Ruangan prescription (CBRP) on rat model with pulmonary fibrosis in duced by bleomycin. Methods: Fifty four male Sprague Dawley rats were rando mly divided into 6 groups (9 rats in each group). From the first day to the 28th day of the experiment, except to those in the sham model control group that were treated with normal saline, the same amount of bleomycin injection as the n ormal saline given to the control group was given through endotracheal instillat ion to all the rats in all the other groups. From the 29th day of the modeling, CBRP solution of different dosages was respectively injected into the rats in th e high, moderate and low CBRP dose group, while equal volume of normal saline w as given to those in the sham model control group and the model control group , and an equal volume of prednisone solution was given to rats in the prednisone group. On the 80th day, the high resolution computerized tomographic (HRCT) images were observed on an equal footing, and HRCT pathology was correlativel y studied. Results: Different HRCT pathological changes were shown in th e rats with pulmonary fibrosis, such as lung consolidation, thickening of interl obular septum and interlobular mesenchyma as well as lobular deformation, nodule shadow, abnormal brochiovascular tract, thickened pleura with irregular junctio n and polished glass like dense shadows. Honeycomb lung was observed in some cases. Pathological sections showed fibrotic proliferation of lung tissues and noticeable pulmonary interstitial fibrosis. CBRP could improve HRCT images of rats with pulmonary fibrosis, and lower fibrotic p roliferation of the lung tissue.Conclusion: CBRP plays its therapeutic role possibly through its effect on the structure of the lung in rats with pulmonary fibrosis.

The protective and therapeutic effects of total flavonoids of Astragalus against bleomycin-induced pulmonary fibrosis are through the enhancement of autophagy

Background:Previously,we showed that total flavonoids from astragalus(TFA)had beneficial effects against transforming growth factor(TGF)-β1-mediated fibrosis,but whether these effects involved autophagy is not known.We attempt to explore the effects of TFA on autophagy in an animal model of idiopathic pulmonary fibrosis(IPF)induced by bleomycin,and to look for TFA components that may have an effect on autophagy.Methods:C57BL/6 mice were randomized to the sham group(SG),model group,low-dose TFA group(LDTG),and high-dose of TFA group(HDTG).The A549 cell line was treated with the TFA components including formononetin,calycosin,isorhamnetin,kaempferol,and quercetin.Lung tissues and cells were examined by histology,immunohistochemistry(anti-TGF-β1,α-smooth muscle actin(SMA),and cadherin),immunofluorescence(microtubule-associated protein light chain 3(LC3)),hydroxyproline content,and immunoblotting(smad3,smad7,phosphoinositide 3-kinase(PI3K),α-SMA,E-cadherin,beclin-1,and LC3-Ⅱ).Results:In vivo,TFA inhibited TGF-β1 expression and decreased collagen content in lung tissues induced by bleomycin.TFA increased autophagy following suppression of the smad pathway.In vitro,quercetin inhibited the epithelial-mesenchymal transition(EMT)of A549 cells induced by TGF-β1 through suppression of the smad pathway.Autophagy was also increased by quercetin through inhibition of the AKT/mTOR pathway,but without change in PI3K expression.Formononetin,calycosin,isorhamnetin,and kaempferol had no such effects.Conclusion:TFA can alleviate bleomycin-induced PF in C57BL/6 mice via enhanced autophagy.The smad and AKT/mTOR pathways are possibly involved in these effects.Quercetin was the main active compound in TFA.

Effect of expression of TGF-beta and TNF-alpha with Qidan granule treatment in rats by bleomycin-induced pulmonary fibrosis

Objective: To evaluate the therapeutic effects and mechanisms of Qidan granule in blemycinA5-induced pulmonary interstitial fibrosis (PIF)in rats. Methods: PIF models were established by blemycinA5-induced in rats. They were treated by Qidan granule and Hydrocortisone respectively. The pathological changes and collagen protein disposition were observed, and the expression of TGF-β, TNF-α proteins were measured by immunohistochemical technique. Results: The pulmonary alveolitis and fibrosis were alleviated remarkably in Qidan granule group compared with those in the model control group and hydrocortisone group (P<0.01). The expression of TGF-β and TNF-α protein were higher in Qidan granule group than those in normal group ,and were significantly less than those in the model control group and in hydrocortisone group (P<0.01). Conclusion: Qidan granule would ameliorate the pulmonary alveolitis and fibrosis. TGF-β and TNF-α might play an important role in the development of alveolitis and fibrosis in rats.

Strengthening pharmacotherapy research for COVID-19-induced pulmonary fibrosis

The global spread of severe acute respiratory syndrome coronavirus 2 has resulted in a significant number of individuals developing pulmonary fibrosis(PF),an irreversible lung injury.This condition can manifest within a short inter-val following the onset of pneumonia symptoms,sometimes even within a few days.While lung transplantation is a potentially lifesaving procedure,its limited availability,high costs,intricate surgeries,and risk of immunological rejection present significant drawbacks.The optimal timing of medication administration for coronavirus disease 2019(COVID-19)-induced PF remains controversial.Despite this,it is crucial to explore pharmacotherapy interventions,involving early and preventative treatment as well as pharmacotherapy options for advanced-stage PF.Additionally,studies have demonstrated disparities in anti-fibrotic treatment based on race and gender factors.Genetic mutations may also impact therapeutic efficacy.Enhancing research efforts on pharmacotherapy interventions,while considering relevant pharmacological factors and optimizing the timing and dosage of medication administration,will lead to enhanced,personalized,and fair treatment for individuals impacted by COVID-19-related PF.These measures are crucial in lessening the burden of the disease on healthcare systems and improving patients'quality of life.

A novel pulmonary fibrosis murine model with immune-related liver injury

Idiopathic pulmonary fibrosis(IPF),characterized by aggravated alveolar destruc-tion and fibrotic matrix deposition,tendentiously experiences the stage called acute exacerbation IPF(AE-IPF)and progresses to multiple organ damage,especially liver injury.Recent studies have found a variety of immune microenvironment disorders associated with elevated IPF risk and secondary organ injury,whereas current animal models induced with bleomycin(BLM)could not completely reflect the pathologi-cal manifestations of AE-IPF patients in clinic,and the exact underlying mechanisms are not yet fully explored.In the current study,we established an AE-IPF model by tracheal administration of a single dose of BLM and then repeated administrations of lipopolysaccharide in mice.This mouse model successfully recapitulated the clinical features of AE-IPF,including excessive intrapulmonary inflammation and fibrosis and extrapulmonary manifestations,as indicated by significant upregulation of Il6,Tnfa,Il1b,Tgfb,fibronectin,and Col1a1 in both lungs and liver and elevated serum aspartate transaminase and alanine transaminase levels.These effects might be attributed to the regulation of Th17 cells.By sharing this novel murine model,we expect to pro-vide an appropriate experimental platform to investigate the pathogenesis of AE-IPF coupled with liver injury and contribute to the discovery and development of targeted interventions.

Identification of potential biomarkers for idiopathic pulmonary fibrosis and validation of TDO2 as a potential therapeutic target

BACKGROUND Idiopathic pulmonary fibrosis(IPF)is a progressive interstitial lung disease with a high mortality rate.On this basis,exploring potential therapeutic targets to meet the unmet needs of IPF patients is important.AIM To explore novel hub genes for IPF therapy.METHODS Here,we used public datasets to identify differentially expressed genes between IPF patients and healthy donors.Potential targets were considered based on multiple bioinformatics analyses,especially the correlation between hub genes and carbon monoxide diffusing capacity of carbon monoxide,forced vital capacity,and patient survival rate.The mRNA levels of the hub genes were determined through quantitative real-time polymerase chain reaction.RESULTS We found that TDO2 was upregulated in IPF patients and predicted poor prognosis.Surprisingly,single-cell RNA sequencing data analysis revealed significant enrichment of TDO2 in alveolar fibroblasts,indicating that TDO2 may participate in the regulation of proliferation and survival.Therefore,we verified the upregulated expression of TDO2 in an experimental mouse model of transforming growth factor-β(TGF-β)-induced pulmonary fibrosis.Furthermore,the results showed that a TDO2 inhibitor effectively suppressed TGF-β-induced fibroblast activation.These findings suggest that TDO2 may be a potential target for IPF treatment.Based on transcription factors-microRNA prediction and scRNA-seq analysis,elevated TDO2 promoted the IPF proliferation of fibroblasts and may be involved in the P53 pathway and aggravate ageing and persistent pulmonary fibrosis.CONCLUSION We provided new target genes prediction and proposed blocking TGF-βproduction as a potential treatment for IPF.

Acute exacerbation of idiopathic pulmonary fibrosis treated using the Feibi recipe:Two case reports

BACKGROUND Rationale:No other treatment besides lung transplant is effective for idiopathic pulmonary fibrosis(IPF).Patients with IPF have poor prognosis,which may eventually lead to death.Patient concerns:Two female patients were diagnosed with IPF.In our recent follow-up,both these patients maintained a good quality of life.CASE SUMMARY Diagnosis:Both patients had dry cough and progressive dyspnea.Interventions:The first patient was treated with prednisone,and the second patient was treated with prednisone and tripterygium glycosides.However,the symptoms did not improve and fibrosis was not controlled.Thus,the Feibi recipe was used.Outcomes:No deterioration was observed after the treatment,and the dry cough and its effect were ameliorated.Furthermore,they are still alive and the quality of their lives has improved.CONCLUSION These two cases suggest that the Feibi recipe and other traditional Chinese medicine therapies could be beneficial for IPF treatment.

The effect of Huayu Lifei formula on the expression of miR-27a and α-SMA in lung tissue of bleomycin-induced rat lung fibrosis model

Objective:Investigating the inhibitory effect of Huayu Lifyei Formula on bleomycininduced rat pulmonary fibrosis and its impact on the expression of miR-27a andα-SMA.Methods:Wistar rats were arbitrarily classified into a normal group,a model group,and a group treated with Huayu Lifyei Formula,each consisting of ten rats.Pulmonary fibrosis rat model was established by injecting bleomycin.Subsequent to the modeling,the Huayu Lifyei Formula treatment group was administered Huayu Lifyei Formula via gavage for a period of 7 days.Rats were sacrificed on the 14th day after modeling.The right lung was taken for HE staining,Masson staining,and immunohistochemical observation of alpha-smooth muscle actin(α-SMA)expression.The expression of miR-27a was measured by qRT-PCR,with the miR-27a binding site on ACTA2(the gene encodingα-SMA protein)confirmed using dualluciferase reporter gene technology.Results:When compared to the model group,the Huayu Lifyei Formula treatment group showed considerable alleviation of pathological morphological changes in lung tissue,with significant reductions in alveolitis,fibrosis,collagen deposition in lung tissue,and the expression ofα-SMA protein.Meanwhile,the expression of miR-27a in the Huayu Lifyei Formula treatment group significantly increased,and the dual-luciferase reporter gene confirmed the binding site of miR-27a with the ACTA2 gene.Conclusion:Huayu Lifyei Formula can inhibit bleomycin-induced pulmonary fibrosis in rats,and its mechanism may be related to the promotion of miR-27a expression.

Inhalation therapy for pulmonary fibrosis:chemical medicines and herbal medicines

Pulmonary fibrosis(PF)is a chronic,progressive,and irreversible pulmonary interstitial disease with unclear pathogenesis.Currently,there are few treatment options for managing PF.Inhalation therapy,as a routine treatment for respiratory diseases,is being used to study the treatment of PF.Some herbal medicines and their active ingredients have been reported to have anti-PF effects.This review aims to provide an overview of the latest developments in inhalation therapy,focusing on the utilization of chemical medicines and herbal medicines for the treatment of PF in both clinical practice and basic research.The inhalation of chemical drugs such as pirfenidone,nintedanib,N-acetylcysteine,and interferon-γhas been shown to demonstrate anti-PF effects.Additionally,the inhalation of various natural products derived from herbal medicines,encompassing polyphenols,alkaloids,flavonoids,saponins,terpenoids,and herbal extracts,contributes to the therapeutic management of PF through diverse mechanisms.The inhalation of both chemical and herbal medicines presents promising advantages in the treatment of PF.Further clinical trials are required to investigate the effectiveness,safety,and mechanism of action of inhalation therapy utilizing natural products derived from herbal medicines.

Jinbei Oral Liquid ameliorates bleomycin-induced idiopathic pulmonary fibrosis in rats via reversion of Th1/Th2 shift

Objective:Idiopathic pulmonary fibrosis(IPF)is a progressive and lethal interstitial lung disease with high mortality.The pivotal role of Th1/Th2 immunological balance in the development and progression of IPF has been demonstrated previously.This study aimed to evaluate the effect of Jinbei Oral Liquid(JBOL)on IPF and its relationship with Th1/Th2 shift.Methods:Rats were divided into six groups:control group,model group(bleomycin),pirfenidone group(positive group,54 mg/kg,i.g.)and JBOL(5.4,10.8 and 21.6 mL/kg,i.g.)groups.The rat model was established by an intratracheal instillation of bleomycin(BLM,5 mg/kg).One day after injection of BLM,pirfenidone or JBOL was given to rats once daily within 28 consecutive days,respectively.Positron emission tomography/computed tomography(PET/CT)was performed on the treated rats.The extent of alveolitis and fibrosis was observed by H&E and Masson trichrome staining.The contents of TGF-β1,TNF-α,IL-4 and IFN-γwere further quantified by ELISA assay.Results:PET/CT and histopathological evidence showed the ability of JBOL to attenuate bleomycininduced alveolitis and fibrosis extent,and the alveolitis lesion score was markedly decreased compared with the model group.The increased expression of inflammatory cytokines TGF-β1 and TNF-αinduced by bleomycin was also suppressed by JBOL.The Th1 response was limited by the reduced IFN-γafter BLM administration,and the Th2 response predominated significantly marked by the increased IL-4.JBOL could increase the level of IFN-γand markedly increased the ratio of IFN-γ/IL-4.Conclusion:These findings suggested that JBOL may attenuate BLM-induced idiopathic pulmonary fibrosis via reducing inflammatory cell infiltration,pro-inflammatory cytokine release and excessive collagen deposition in rats.One of the mechanisms is the reversion of Th1/Th2 shift caused by BLM.

Drug repurposing of histone deacetylase inhibitors that alleviate neutrophilic inflammation in acute lung injury and idiopathic pulmonary fibrosis via inhibiting leukotriene a4 hydrolase and blocking LTB4 biosynthesis

OBJECTIVE Leukotriene B4(LTB4)biosynthesis and subsequently neutrophilic inflammation may provide a potential strategy for the treatment of acute lung injury(ALI)or idiopathic pulmonary fibrosis(IPF).To provide a potential strategy for the treatment of ALI or IPF,we identified potent inhibitors of Leukotriene A4 hydrolase(LTA4H),a key enzyme in the biosynthesis of LTB4.METHODS In this study,we identified two known histone deacetylase(HDAC)inhibitors,suberanilohydroxamic acid(SAHA)and its analogue 4-(dimethylamino)-N-[7-(hydroxyamino)-7-oxoheptyl]benzamide(M344),as effective inhibitors of LTA4H using enzymatic assay,thermofluor assay,and X-ray crystallographic investigation.We next tested the effect of SAHA and M344 on endogenous LTB4 biosynthesis in neutrophils by ELISA and neutrophil migration by transwell migration assay.A murine experimental model of ALI was induced by lipopolysaccharide(LPS)inhalation.Histopathological analysis of lung tissue using H&E staining revealed the serious pulmonary damage caused by LPS treatment and the effect of the SAHA.We next examined m RNA and protein levels of pro-inflammatory cytokines in lung tissue and bronchoalveolar lavage fluid using q RT-PCR and ELISA to further investigate the underlying mechanisms of anti-inflammatory activities by SAHA.We also investigated the effects of SAHA and M344 on a murine experimental model of bleomycin(BLM)-induced IPF model.RESULTS The results of enzymatic assay and X-ray crystallography showed that both SAHA and M344 bind to LTA4H,significantly decrease LTB4 levels in neutrophil,and markedly diminish early neutrophilic inflammation in mouse models of ALI and IPF under a clinical safety dose.CONCLUSION Collectively,SAHA and M344 would provide promising agents with well-known clinical safety for potential treatment in patients with ALI and IPF via pharmacologically inhibiting LAT4H and blocking LTB4 biosynthesis.

Salvianolic acid B dry powder inhaler for the treatment of idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis(IPF)is a serious and fatal pulmonary inflammatory disease with an increasing incidenceworldwide.The drugs nintedanib and pirfenidone,are listed as conditionally recommended drugs in the“Evidence-Based Guidelines for the Diagnosis and Treatment of Idiopathic Pulmonary Fibrosis”.However,these two drugs have many adverse reactions in clinical application.Salvianolic acid B(Sal B),a water-soluble component of Salvia miltiorrhiza,could alleviate bleomycin-induced peroxidative stress damage,and prevent or delay the onset of IPF by regulating inflammatory factors and fibrotic cytokines during the disease’s progression.However,Sal B is poorly absorbed orally,and patient compliance is poor when administered intravenously.Therefore,there is an urgent need to find a new non-injection route of drug delivery.In this study,Sal B was used as model drug and l-leucine(LL)as excipient to prepare Sal B dry powder inhaler(Sal B-DPI)by spray drying method.Modern preparation evaluation methods were used to assess the quality of Sal B-DPI.Sal B-DPI is promising for the treatment of IPF,according to studies on pulmonary irritation evaluation,in vivo and in vitro pharmacodynamics,metabolomics,pharmacokinetics,and lung tissue distribution.

Differential Mechanisms of the Effect of Peroxisome Proliferator-Activated Receptor Gamma Agonists on Bleomycin-Induced Lung Fibrosis

Background and Objectives: Peroxisome proliferator-activated receptor-g (PPAR-g) is a nuclear receptor whose activation regulates inflammation and fibrosis in various organs. We aimed to investigate the effect of two PPAR-g ligands, telmisartan and rosiglitazone, on lung injury and fibrosis induced by intratracheal bleomycin (BLM). Methods: Lung injury and fibrosis was induced in female C57Bl/6 mice by intratracheal instillation of 1.0 mg/kg of BLM. Some of the animals received rosiglitazone intraperitoneally or telmisartan in drinking water. Bronchoalveolar lavage (BAL) was performed 2, 7, 14 or 21 days after BLM instillation for cell counting and measurement of mediators in the lung. In a separate series, the lungs were sampled for collagen assay and histopathological evaluation. Results: Treatment with rosiglitazone or telmisartan significantly attenuated the BLM-induced increases in lung collagen content, pathological score, and inflammatory cells in BAL fluid. Rosiglitazone significantly suppressed BLM-induced elevation of TGF-b1, MCP-1, and IL-6 levels in the lung. In contrast, telmisartan made no changes in these cytokines, whereas it mitigated the BLM-induced increase in prostaglandin F2a in the lung. Higher concentrations of rosiglitazone and telmisartan attenuated proliferation of lung fibroblasts in vitro. Conclusions: Two PPAR-g ligands, rosiglitazone and telmisartan, exert protective effects on BLM-induced lung fibrosis through the suppression of different profibrotic mediators.

Rapamycin attenuates bleomycin-induced pulmonary fibrosis in rats and the expression of metalloproteinase-9 and tissue inhibitors of metalloproteinase-1 in lung tissue

Background Idiopathic pulmonary fibrosis （IPF） is the most common and devastating form of interstitial lung disease （ILD） in the clinic.There is no effective therapy except for lung transplantation.Rapamycin is an immunosuppressive drug with potent antifibrotic activity.The purpose of this study was to examine the effects of rapamycin on bleomycininduced pulmonary fibrosis in rats and the relation to the expression of metalloproteinase-9 （MMP-9） and tissue inhibitor of metalloproteinase-1 （TIMP-1）.Methods Sprague-Dawley rats were treated with intratracheal injection of 0.3 ml of bleomycin （5 mg/kg） in sterile 0.9％ saline to make the pulmonary fibrosis model.Rapamycin was given at a dose of 0.5 mg/kg per gavage,beginning one day before bleomycin instillation and once daily until animal sacrifice.Ten rats in each group were sacrificed at 3,7,14,28 and 56 days after bleomycin administration.Alveolitis and pulmonary fibrosis were semi-quantitatively assessed after HE staining and Masson staining under an Olympus BX40 microscope with an IDA-2000 Image Analysis System.Type Ⅰ and Ⅲ collagen fibers were identified by Picro-sirius-polarization.Hydroxyproline content in lung tissue was quantified by a colorimetric-based spectrophotometric assay,MMP-9 and TIMP-1 were detected by immunohistochemistry and by realtime quantitative reverse transcriptase polymerase chain reaction （RT-PCR）.Results Bleomycin induced alveolitis and pulmonary fibrosis of rats was inhibited by rapamycin.Significant inhibition of alveolitis and hydroxyproline product were demonstrated when daily administration of rapamycin lasted for at least 14 days.The inhibitory efficacy on pulmonary fibrosis was unremarkable until rapamycin treatment lasted for at least 28 days （P ＜0.05）.It was also demonstrated that rapamycin treatment reduced the expression of MMP-9 and TIMP-1 in lung tissue that was increased by bleomycin.Conclusion These results highlight the significance of rapamycin in alleviating alveolitis and pulmonary fibrosis,which is associated with decreased expression of MMP-9 and TIMP-1.

Correlation Analysis between Idiopathic Pulmonary Fibrosis and Tumor Markers

The purpose of this study was to investigate the correlation between idiopathic pulmonary fibrosis(IPF)and tumor markers to provide evidence for early screening of precancerous lesions.In our hospital from July 2017 to May 2019,40 patients with IPF treatment were selected as the IPF group,and 40 patients with idiopathic pulmonary fibrosis with lung cancer(IPF-LC)were selected as the IPF-LC group.In the same period,40 healthy physical examinees were used as control group.Different types of patients in the IPF-LC group were divided into lung adenocarcinoma group,small cell lung cancer group and l squamous carcinoma group.The expression levels of tumor markers were detected in the three groups,the positive rates of tumor markers in IPF group,IPF-LC group and their subgroups were compared.The results showed that the levels of neuron specific enolase(NSE),antigen CYFRA211,carcinogenic antigen(CEA)and cancer antigen 125(CA125)in IPF and IPF-LC groups were significantly higher than those in control group(P<0.05).There was no significant difference in CEA and CYFRA211 between IPF-LC group and IPF group.The level of NSE in IPF-LC group was significantly higher than that in IPF group,while the level of CA125 was significantly lower than that in IPF group(P<0.0.5).The difference of positive rate of NES and CA125 in IPF-LC group and IFP group was statistically significant(P<0.05),there was no statistically significant difference in the positive rate of other indicators(P>0.05)The NSE positive rate of IPF group was significantly lower than that of IPF-LC group(P<0.05),the positive rates of other tumor markers were significantly lower than those of each subgroup of IPF-LC group(P<0.05).Therefore,tumor markers in IPF patients showed different degrees of increase,which is worthy of clinical attention.Among them,NSE can be used as an early screening indicator for IPF precancerous lesions.

Serum biomarkers in evaluation and management of idiopathic pulmonary fibrosis

Despite there has greater understanding of the pathophysiology in idiopathic pulmonary fibrosis,idiopathic pulmonary fibrosis still remains a challenge in clinical practice and translational research due to its heterogeneity.Improvements in molecular techniques have identified potential pathways and targets for diagnosis and therapeutic intervention.Several types of idiopathic pulmonary fibrosis biomarkers such as KL-6,SP-A,SP-D,MMP7 and other potential ones have been studied extensively.In addition,many studies have confirmed that levels of some tumor markers such as CA 19-9,CA 15-3,CEA,CA 125,CYFRA 21-1 are useful for idiopathic pulmonary fibrosis.The present study focuses on serum biomarkers including tumor markers to provide an overview on the current roles of these biomarkers in the setting of diagnosis,prediction as well as response to therapy in idiopathic pulmonary fibrosis.